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1.
目的:广西眼镜王蛇毒一种碱性蛋白的分离纯化和性质研究.方法:分离纯化采用DEAE-Sepharose CL-6B离子交换柱层析,Sephadex G-75凝胶过滤和CM-Sepharose CL-6B离子交换柱层析;通过电泳,磷脂酶A2(PLA2)酶活力测定,氨基酸组成分析,N端序列测定及药理实验进行性质研究.结果:从广西眼镜王蛇毒中分离到一种电泳纯的碱性蛋白(命名为CM-IV),相对分子质量约为14 000,等电点约为8.65;PLA2的比活力为23 μmol·mg-1/min.氨基酸组成分析表明,其分子中Cys的含量占6.85%;利用蛋白质直接测序N端序列为TKCYVTPDVK.药理实验表明它对实验用小白鼠有致死性(LD50约为0.2 mg/kg),并具有心脏毒性,但无血小板抑制活性.根据Genebank Blast 分析软件,它的N端序列与神经毒素具有高度的同源性.结论:这种碱性蛋白可能为神经毒素,但却拥有PLA2的活力.  相似文献   

2.
目的:探索从中华眼镜蛇蛇毒中快速分离提纯磷脂酶A2(PLA2)的方法方法:应用ARG-CY仿生配基,用亲和层析一步法从中华眼镜蛇蛇毒粗毒中纯化PLA2,用SDS-PAGE进行纯度鉴定及相对分子质量测定,并测定其溶血活性.结果:纯化后获得的PLA2电泳图谱呈单一区带,相对分子质量为14 000,活性为182 μmol/(min·mg).产物具有溶血活性.结论:仿生亲和层析法提纯LPA2操作简单,纯度高,为一高效实用的方法.  相似文献   

3.
广西眼镜王蛇(Ophiophagus hannah)蛇毒,经羧甲基纤维素离子交换层析,可得20个蛋白峰,磷脂酶A_2活性以第一峰活性最强,第二和第十二峰也有部分活性.取第一峰观察其对红细胞电泳速度的影响及对红细胞膜影响的电镜观察,发现磷脂酶A_2对红细胞电泳速度有明显的延缓作用,并对人的红细胞膜有明显的作用,但对山羊红细胞膜未见明显改变.研究还表明眼镜王蛇毒中磷脂酶A_2也具有很好的热稳定性.  相似文献   

4.
[目的]鉴定中华眼镜蛇(Naja naja atra)蛇毒中的神经保护因子,研究其抗神经元凋亡作用机制。[方法]连续柱层析法分离、纯化蛇毒神经保护因子。用Edman N-末端测序法取得蛋白质一级序列,BLAST软件进行蛋白质鉴定。使用低钾诱导体外培养小脑颗粒神经元凋亡模型,研究其神经元保护作用。[结果]从中华眼镜蛇蛇毒中分离出一种,可浓度依赖性地抑制低钾诱导的小脑颗粒神经元凋亡。神经保护因子(Cobra venom neu-ronal protective factor,CVNPF)此神经保护因子N-末端20个氨基酸序列为NLYQFKNMIQCTVPSRSWWD-,BLAST序列同源比对确定该保护因子为中华眼镜蛇分泌型磷脂酶A2(secreted phospholipase A2,sPLA2)。进一步发现来源于蜂毒、莫桑比克眼镜蛇(Njaja naja mossambica)蛇毒、西部菱斑响尾蛇(Crotalus atroxalso)蛇毒的sPLA2也对小脑颗粒神经元具有保护作用;其作用不为磷脂酶A2酶活性抑制剂7,7-Dimethyleicosadienoic acid(DEDA)和Manoalide阻断。[结论]CVNPF属于sPLA2。多种sPLA2可以抗神经元凋亡,其保护机制与磷脂酶A2的酶活性无关。  相似文献   

5.
分泌型磷脂酶 A2非酶活性依赖的神经元保护作用   总被引:1,自引:0,他引:1  
【目的】鉴定中华眼镜蛇(Najanajaatra)蛇毒中的神经保护因子,研究其抗神经元凋亡作用机制。【方法】连续柱层析法分离、纯化蛇毒神经保护因子。用EdmanN-末端测序法取得蛋白质一级序列,BLAST软件进行蛋白质鉴定。使用低钾诱导体外培养小脑颗粒神经元凋亡模型,研究其神经元保护作用。【结果】从中华眼镜蛇蛇毒中分离出一种,可浓度依赖性地抑制低钾诱导的小脑颗粒神经元凋亡。神经保护因子(Cobravenomneu-ronalprotectivefactor,CVNPF)此神经保护因子N-末端20个氨基酸序列为NLYQFKNMIQCTVPSRSWWD-,BLAST序列同源比对确定该保护因子为中华眼镜蛇分泌型磷脂酶A2(secretedphospholipaseA2,sPLA2)。进一步发现来源于蜂毒、莫桑比克眼镜蛇(Njajanajamossambica)蛇毒、西部菱斑响尾蛇(Crotalusatroxalso)蛇毒的sPLA2也对小脑颗粒神经元具有保护作用;其作用不为磷脂酶A2酶活性抑制剂7,7-Dimethyleicosadienoicacid(DEDA)和Manoalide阻断。【结论】CVNPF属于sPLA2。多种sPLA2可以抗神经元凋亡,其保护机制与磷脂酶A2的酶活性无关。  相似文献   

6.
目的:从广西眼镜蛇毒中分离出Natrin蛋白.方法:广西眼镜蛇毒经SephadexG75凝胶、CM Sepharose CL-6B离子交换层析及FPLC mono-S柱层析分离纯化出Natrin蛋白.结果:从广西眼镜蛇蛇毒中分离纯化得到的Natrin蛋白,经SDS-PAGE电泳测定其分子量为25 ku,质谱测定其分子量...  相似文献   

7.
目的:研究蛇毒Ⅱ类磷脂酶A2(PLA2)中D49 PLA2和K49 PLA2的功能分化及其功能分化决定位点的鉴定。方法:运用序列比较分析,进化树构建和DIVERGE v1.04软件计算研究D49 PLA2和K49 PLA2的功能分化情况及其分化位点。结果:序列比较分析,进化树构建和DIVERGE v1.04软件计算结果表明蛇毒Ⅱ类PLA2中D49 PLA2和K49 PLA2的确发生了功能分化,对于K49 PLA2来说,1S,7K,11Q,E12,R34,T56,N88,L92,E108,K116,K128可能为功能分化决定位点。对于D49 PLA2,L2,G33,G35,F46和Y118可能为功能分化决定位点。结论:我们首次通过序列比较分析,进化树构建和DIVERGE v1.04软件计算鉴定出蛇毒Ⅱ类PLA2中D49 PLA2和K49 PLA2可能的功能分化位点,为今后通过基因重组和定点突变方法研究蛇毒Ⅱ类PLA2结构功能关系提供了线索。  相似文献   

8.
论 著·广西眼镜王蛇毒酸性磷脂酶 A2 的分离纯化和性质研究 1:1………………………………………………………………………羧乙基锗倍半氧化物对大鼠离体颈上神经节烟碱传递的影响 1:4…………………………………………………………………儿童急性肾炎初期左室舒张功能损害的临床研究 1:7………………………………………………………………………………CD3 AK细胞治疗晚期恶性肿瘤疗效观察 1:10………………………………………………………………………………………鲨鱼软骨提取物对小鼠实体瘤抑制作用的实验研究 1:12…………………………  相似文献   

9.
磷脂酶A2抗菌作用的研究进展   总被引:3,自引:0,他引:3  
梁宁生  刘滔滔 《医学文选》1999,18(5):820-822
磷脂酶A2(PhospholipaseA2,EC3.1.1.4),简称PLA2,是一种重要代谢和调节酶类,它广泛的存在于哺乳动物的细胞和体液以及蛇毒、蜂毒中。从1900年研究胰液和蛇毒中的PLA2开始到现在,已近百年[1],但它们在生理过程中的一些重要作用直到近二十年才逐渐被认识。随着研究的不断深入,新的种类、新的作用不断被发现,使PLA2的研究越来越受到人们的关注。近几年来的一个重要发现是PLA2在宿主防御细菌感染中的作用-直接的杀菌作用[2,3]。1 PLA2的类型和一般作用  目前发现和提纯的PLA2已超过100种[4],根据PLA2氨基酸数目、序列、结构特点和…  相似文献   

10.
福建产舟山眼镜蛇毒细胞毒素的快速分离纯化及鉴定   总被引:8,自引:4,他引:4  
目的 从舟山眼镜蛇毒中分离纯化细胞毒素(CTX),并鉴定其理化性质。方法 采用SP—Sephadex C-25阳离子交换色谱及Sephasil Peptide C18反相高效液相色谱法从舟山眼镜蛇毒中分离纯化CTX,SDS—PAGE(Tris—Tricine系统)鉴定纯度,Edman降解法测定N端氨基酸序列。结果 粗毒经阳离子交换色谱,得到14个蛋白峰,其中第X~XⅢ峰具CTX活性;再分别经反相色谱纯化,得到4个CTX.总得率为32.48%;SDS—PAGE显示为均一蛋白,分子量依次为:7.28,7.33,7.24和7.38kD;测定它们N端20个氨基酸序列。结论 采用阳离子交换和反相高效液相色谱可快速、高效地从眼镜蛇毒中获得4个CTX纯品。  相似文献   

11.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

12.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

13.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

14.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

15.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

16.
A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care(~'4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.  相似文献   

17.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

18.
Journal of Nanjing Medical University (English Edition) JNMU, sponsored by Nanjing Medical University, was established in 1987. It is a bimonthly comprehensive English medical journal published locally and abroad.Since 2007, Journal of Nanjing Medical University (English Edition )was granted Elsevier the full publishing and distribution rights worldwide for the Electronic Edition, excluding the People's Republic of China.  相似文献   

19.
Objective To measure Derpl and Blot5 allergen levels in asthmatics' homes in Hongkong.Methods Seventy houses were enrolled for a mite indoor environment study. Dust samples were obtained from two sites of each patients' house: bed and floor. Derpl and Blot5 levels were quantified by a two-site monoclonal antibody-based ELISA technique.Results The levels of Derpl allergens found in bed (geometric mean (GM) 3.43 μg/g of dust; 95%CI, 1.89-4.96 μg/g)and on the floor (GM 1.12 μg/g of dust; 95%CI, 0.71-1.53 μg/g) indicated significant differences (P=0.005). However, the levels of Blot5 allergens found in bed (GM 19.00 μg/g of dust; 95%CI, 0.89-38.90 μg/g) and on the floor (GM 6.14 μg/g of dust; 95%CI, 0.40-11.90 μg/g) showed no statistically significant difference. In addition, in regards to the exposure index for Derpl and Blot5 allergens found in bed and on the floor, 17.6% in bed and 8.6% on the floor had levels of Blot5 ≥ 10 μg/g of dust, higher than those obtained for Derp1 (7.2% and 0% in bed and on the floor respectively, P< 0.05); higher percentages in bed and on the floor (25.0% and 35.7%) were observed for levels of Blot5 =0 μg/g of dust as compared with Derpl in bed and on the floor (4.3% and 14.5% respectively, P< 0.05).Conclusions Derpl and Blot5 are the major allergens found in this regional study, Blot5 is a more potent allergen in Hongkong, probably reflecting the high level of exposure to Blomia tropicalis (Bt). Bt and Dermatophagoides pteronyssinus (Dp) allergens should be included for precise diagnosis and effective immuno-therapeutic treatment of mite allergy in Hongkong.  相似文献   

20.
ANTI-PROLIFERATION EFFECT OF ORIDONIN ON HL-60 CELLS AND ITS MECHANISM   总被引:4,自引:0,他引:4  
Objcetive To investigate the anti-proliferation effect of oridonin on leukemic HL-60 cells and its mechanism. Methods HL-60 cells in vitro in culture medium were given different concentrations of oridonin. The inhibitory rate of cells were measured by microculture tetrazolium (MTT) assay, cell apoptotic rate was detected by flow cytometry (FCM), morphology of cell apoptosis was observed by hoechst 33258 fluorescence staining, and the activity of telomerase was detected using telomere repeat amplification protocol (TRAP) PCR-ELISA before and after apoptosis occurred. Results Oridonin could decrease telomerase activity, inhibit growth of HL-60 cells, and cause apoptosis significantly. The suppression was both in time- and dose-dependent manner. Marked morphological changes of cell apoptosis including condensation of chromatin and nuclear fragmentation were observed clearly by hoechst 33258 fluorescence staining especially after cells were treated 48-60 hours by oridonin. Conclusions Oridonin has apparent anti-proliferation and apoptotic effects on HL-60 cells in vitro, decreasing telomerase activity of HL-60 cells may be one of its most important mechanisms. These results will provide strong laboratory evidence of oridonin for clinical treatment of acute leukemia.  相似文献   

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